Pharmaceuticals in the Aquatic Environment: No Answers Yet to the Major Questions

Data Availability Statement: Data are not used in this manuscript.Copyright © 2022 The Authors. The presence of pharmaceuticals in the environment, especially the aquatic environment, has received a lot of attention in the last 20 plus years. Despite that attention, the two most important questions regarding pharmaceuticals in the environment still cannot be answered. It is not possible to put the threat posed by pharmaceuticals into perspective with the many other threats (stressors) facing aquatic organisms, such as low flows due to over-abstraction of water, inhibited passage of migratory species due to dams and weirs, diseases, algal blooms causing low oxygen levels and releasing toxins, eutrophication, climate change, and so on. Nor is it possible to identify which pharmaceuticals are of concern and which are not. Not only can these key questions not be answered presently, they have received extremely little attention, despite being identified 10 years ago as the two most important questions to answer. That situation must change if resources and expertise are to be effectively used to protect the environment.Natural Environment Research Council. Grant Number: NE/S000100/1 CHEMPOP

The consequences of feminization in breeding groups of wild fish

EHP is a publication of the U.S. government. Publication of EHP lies in the public domain and is therefore without copyright. Research articles from EHP may be used freely; however, articles from the News section of EHP may contain photographs or figures copyrighted by other commercial organizations and individuals that may not be used without obtaining prior approval from both the EHP editors and the holder of the copyright. Use of any materials published in EHP should be acknowledged (for example, "Reproduced with permission from Environmental Health Perspectives") and a reference provided for the article from which the material was reproduced.BACKGROUND: The feminization of nature by endocrine-disrupting chemicals (EDCs) is a key environmental issue affecting both terrestrial and aquatic wildlife. A crucial and as yet unanswered question is whether EDCs have adverse impacts on the sustainability of wildlife populations. There is widespread concern that intersex fish are reproductively compromised, with potential population-level consequences. However, to date, only in vitro sperm quality data are available in support of this hypothesis. OBJECTIVE: The aim of this study was to examine whether wild endocrine-disrupted fish can compete successfully in a realistic breeding scenario. METHODS: In two competitive breeding experiments using wild roach (Rutilus rutilus), we used DNA microsatellites to assign parentage and thus determine reproductive success of the adults. RESULTS: In both studies, the majority of intersex fish were able to breed, albeit with varying degrees of success. In the first study, where most intersex fish were only mildly feminized, body length was the only factor correlated with reproductive success. In the second study, which included a higher number of more severely intersex fish, reproductive performance was negatively correlated with severity of intersex. The intersex condition reduced reproductive performance by up to 76% for the most feminized individuals in this study, demonstrating a significant adverse effect of intersex on reproductive performance. CONCLUSION: Feminization of male fish is likely to be an important determinant of reproductive performance in rivers where there is a high prevalence of moderately to severely feminized males.Funding for this work was derived through the Endocrine Disruption in Catchments project, which was supported by the U.K. Department for Environment Food and Rural Affairs and the U.K. Environment Agency

A comprehensive aquatic risk assessment of the beta-blocker propranolol, based on the results of over 600 research papers

A comprehensive aquatic environmental risk assessment (ERA) of the human pharmaceutical propranolol was conducted, based on all available scientific literature. Over 200 papers provided information on environmental concentrations (77 of which provided river concentrations) and 98 dealt with potential environmental effects. The median concentration of propranolol in rivers was 7.1 ng/L (range of median values of individual studies 0.07 to 89 ng/L), and the highest individual value was 590 ng/L. Sixty-eight EC50 values for 35 species were available. The lowest EC50 value was 0.084 mg/L. A species sensitivity distribution (SSD) provided an HC50 value of 6.64 mg/L and an HC5 value of 0.22 mg/L. Thus, there was a difference of nearly 6 orders of magnitude between the median river concentration and the HC50 value, and over 4 orders of magnitude between the median river concentration and the HC5 value. Even if an assessment factor of 100 was applied to the HC5 value, to provide considerable protection to all species, the safety margin is over 100-fold. However, nearly half of all papers reporting effects of propranolol did not provide an EC50 value. Some reported that very low concentrations of propranolol caused effects. The lowest concentration reported to cause an effect - in fact, a range of biochemical and physiological effects on mussels - was 0.3 ng/L. In none of these ‘low concentration’ papers was a sigmoidal concentration-response relationship obtained. Although inclusion of data from these papers in the ERA cause a change in the conclusion reached, we are sceptical of the repeatability of these ‘low concentration’ results. We conclude that concentrations of propranolol present currently in rivers throughout the world do not constitute a risk to aquatic organisms. We discuss the need to improve the quality of ecotoxicology research so that more robust ERAs acceptable to all stakeholders can be completed.AstraZeneca; Natural Environment Research Counci

Protecting the environment from psychoactive drugs: Problems for regulators illustrated by the possible effects of tramadol on fish behaviour

© 2019 The Authors. There is concern that psychoactive drugs present in the aquatic environment could affect the behaviour of fish, and other organisms, adversely. There is considerable experimental support for this concern, although the literature is not consistent. To investigate why, fish were exposed to three concentrations of the synthetic opiate tramadol for 23–24 days, and their anxiolytic behaviour in a novel tank diving test was assessed both before and after exposure. The results were difficult to interpret. The positive control drug, the anti-depressant fluoxetine, produced the expected results: exposed fish explored the novel tank more, and swam more slowly while doing so. An initial statistical analysis of the results provided relatively weak support for the conclusion that both the low and high concentrations of tramadol affected fish behaviour, but no evidence that the intermediate concentration did. To gain further insight, UK and Japanese experts in ecotoxicology were asked for their independent opinions on the data for tramadol. These were highly valuable. For example, about half the experts replied that a low concentration of a chemical can cause effects that higher concentrations do not, although a similar number did not believe this was possible. Based both on the inconclusive effects of tramadol on the behaviour of the fish and the very varied opinions of experts on the correct interpretation of those inconclusive data, it is obvious that more research on the behavioural effects of tramadol, and probably all other psychoactive drugs, on aquatic organisms is required before any meaningful risk assessments can be conducted. The relevance of these findings may apply much more widely than just the environmental risk assessment of psychoactive drugs. They suggest that much more rigorous training of research scientists and regulators is probably required if consensus decisions are to be reached that adequately protect the environment from chemicals.Ecotoxicology Research Group, Brunel University London funded the fish experiments. We would also like to thank Dr. Matt Winter, University of Exeter, for his support with the behavioural analysis. This study was also supported by the Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT) to a project on Joint Usage/Research Centre – Leading Academia in Marine and Environment Pollution Research (LaMer), and Japan Society for the Promotion of Science (JSPS) Grants-in-Aid (KAKENHI) for JSPS Fellows (JP26·2800), Scientific Research (A) (JP25257403), Scientific Research (A) (JP16H01784), and Young Scientists (JP18K18206)

Quantitative cross-species extrapolation between humans and fish: The case of the anti-depressant fluoxetine

This article has been made available through the Brunel Open Access Publishing Fund.Fish are an important model for the pharmacological and toxicological characterization of human pharmaceuticals in drug discovery, drug safety assessment and environmental toxicology. However, do fish respond to pharmaceuticals as humans do? To address this question, we provide a novel quantitative cross-species extrapolation approach (qCSE) based on the hypothesis that similar plasma concentrations of pharmaceuticals cause comparable target-mediated effects in both humans and fish at similar level of biological organization (Read-Across Hypothesis). To validate this hypothesis, the behavioural effects of the anti-depressant drug fluoxetine on the fish model fathead minnow (Pimephales promelas) were used as test case. Fish were exposed for 28 days to a range of measured water concentrations of fluoxetine (0.1, 1.0, 8.0, 16, 32, 64 μg/L) to produce plasma concentrations below, equal and above the range of Human Therapeutic Plasma Concentrations (HTPCs). Fluoxetine and its metabolite, norfluoxetine, were quantified in the plasma of individual fish and linked to behavioural anxiety-related endpoints. The minimum drug plasma concentrations that elicited anxiolytic responses in fish were above the upper value of the HTPC range, whereas no effects were observed at plasma concentrations below the HTPCs. In vivo metabolism of fluoxetine in humans and fish was similar, and displayed bi-phasic concentration-dependent kinetics driven by the auto-inhibitory dynamics and saturation of the enzymes that convert fluoxetine into norfluoxetine. The sensitivity of fish to fluoxetine was not so dissimilar from that of patients affected by general anxiety disorders. These results represent the first direct evidence of measured internal dose response effect of a pharmaceutical in fish, hence validating the Read-Across hypothesis applied to fluoxetine. Overall, this study demonstrates that the qCSE approach, anchored to internal drug concentrations, is a powerful tool to guide the assessment of the sensitivity of fish to pharmaceuticals, and strengthens the translational power of the cross-species extrapolation

Renewing and improving the environmental risk assessment of chemicals

The processes underpinning the environmental risk assessment (ERA) of chemicals have not changed appreciably in the last 30 years. It is unclear how successful these processes are in protecting the environment from any adverse effects of chemicals. To ascertain if the current methodology can be improved, and if so, how, we invited experts to suggest how the current ERA process could be improved. They were not asked to select from a list of suggestions. The 36 experts made 109 suggestions for improvement, which could be grouped into 33 categories. The category that received the most support, from 12 experts, was to utilise a broader range of scientific information, including all up-to-date information, in ERAs. The second most popular category, supported by 10 experts, was the suggestion to regulate mixtures of chemicals; the current regulatory process involves chemical-by-chemical assessment. Two quite radical proposals were suggested. One was to replace the regulator with artificial intelligence. The other was to establish a new competent authority that would appoint groups of experts, each including representatives of the range of stakeholders, to decide which studies were required, commission those studies, then conduct the ERA based on the results of those studies. These two radical proposals, which the authors support strongly, are not necessarily mutually exclusive. We conclude that the present ERA process could be improved to better protect the environment from the myriad of chemicals in use

A ‘Limitations’ section should be mandatory in all scientific papers

Data availability: No data was used for the research described in the article.NERC for grant NE/S000100/1 supporting the ChemPop project on the impacts of chemicals on wildlife

The consequences of exposure to mixtures of chemicals: Something from ‘nothing’ and ‘a lot from a little’ when fish are exposed to steroid hormones

Ill-defined, multi-component mixtures of steroidal pharmaceuticals are present in the aquatic environment. Fish are extremely sensitive to some of these steroids. It is important to know how fish respond to these mixtures, and from that knowledge develop methodology that enables accurate prediction of those responses. To provide some of the data required to reach this objective, pairs of fish were first exposed to five different synthetic steroidal pharmaceuticals (one estrogen, EE2; one androgen, trenbolone; one glucocorticoid, beclomethasone dipropionate; and two progestogens, desogestrel and levonorgestrel) and concentration-response data on egg production obtained. Based on those concentration-response relationships, a five component mixture was designed and tested twice. Very similar effects were observed in the two experiments. The mixture inhibited egg production in an additive manner predicted better by the model of Independent Action than that of Concentration Addition. Our data provide a reference case for independent action in an in vivo model. A significant combined effect was observed when each steroidal pharmaceutical in the mixture was present at a concentration which on its own would produce no statistically significant effect (something from ‘nothing’). Further, when each component was present in the mixture at a concentration expected to inhibit egg production by between 18% (Beclomethasone diproprionate) and 40% (trenbolone), this mixture almost completely inhibited egg production: a phenomenon we term ‘a lot from a little’. The results from this proof-of-principle study suggest that multiple steroids present in the aquatic environment can be analysed for their potential combined environmental risk